Process and system for obtaining botulinum neurotoxin

摘要

Rapid, animal protein free, chromatographic processes and systems for obtaining high potency, high yield botulinum neurotoxin for research, therapeutic and cosmetic use.

主权项

1. A process for producing a biologically-active botulinum neurotoxin type A complex, the process comprising the following steps:
(a) providing a substantially APF fermentation medium; (b) fermenting Clostridium botulinum bacteria in the fermentation medium, wherein the Clostridium botulinum bacteria produces a botulinum neurotoxin, and; (c) harvesting the fermentation medium by removing cellular debris; (d) concentrating the harvested fermentation medium by filtration; (e) diluting the concentrated fermentation medium by adding a buffer thereto; (f) performing a first contacting step in which the diluted, harvested fermentation medium is contacted with an anion exchange chromatography medium so that biologically-active botulinum neurotoxin in the medium is captured by the anion exchange medium; (g) eluting captured botulinum neurotoxin from the anion exchange medium to obtain a first eluate; (h) performing a second contacting step in which the first eluate is contacted with a cation exchange chromatography medium to remove impurities from the first eluate, thereby obtaining a second eluate; (i) performing a third contacting step in which the second eluate is contacted with a hydrophobic interaction chromatography medium and eluted to obtain a third eluate; (j) processing the third eluate by diafiltration; and (k) filtering the processed third eluate to obtain a biologically-active botulinum neurotoxin type A complex-containing product having a potency of 2.0×107 to 6.0×107 units/mg.